Ankle injuries are amongst the most common musculoskeletal injuries. The necessity of prevention measurements before or an early rehabilitation start after an injury, is essential for (semi-) professional sports like soccer to decrease healing duration. Sensor-supported serious games could complement a therapeutic program to support resilience and motivation during the prevention or rehabilitation process. Therefore, the aim of this study is to develop and evaluate a user-centered prototype of a serious game using a commercial Off-The-Shelf MetaMotion IMU sensor. A semi-structured interview with a soccer club therapist, followed by an online questionnaire containing 48 questions (n = 91), was performed to ensure a user-centered approach. Based on this, a prototype, including five identified functional requirements and seven exercises (comprising: horizontal/vertical in- and eversion, dorsi- and plantarflexion, knee bend and squat, and toe and heel rise), was developed in an iterative process and evaluated by two participants with an acute ankle injury. The questionnaire outcomes showed averages of 3.3 ankle injuries per participant and 40 absence days per incident. Additionally, 85% of the participants reported needing more prevention time for such injuries. The evaluation phase (total training duration: 2 h 52 min) consisted of playing two different game types (1 and 2 degrees of freedom) and three different levels, where an avatar needs to be controlled while running and avoiding obstacles or collecting trophies. Both range of motion (ROM) and scores, which are directly measured by the game, showed significant improvements (ROM: t = 5.71; p < 0.01; Score: t = 3.98; p < 0.01) between the first and last session in both participants (P1: ROM +3.56°; Score +7.00%, P2: ROM +6.59°; Score +9.53%), indicating high effectiveness, despite a short training period (1 and 2 weeks). ROM improvement results and athlete feedback coincide in that the sensor-assisted serious game might be beneficial for ankle prevention and rehabilitation. At the same time, the increased scores indicate substantial motivation over several training sessions.